The parvoviruses AMDV and B19 are unique among animal DNA viruses in that they have a single promoter, and so their genetic diversity is controlled exclusively by post-transcriptional mechanisms. A single class of pre-mRNA molecules generated by these viruses undergoes extensive alternative splicing and polyadenylation that generates sub-genomic mRNA molecules which program this diversity. Control of the synthesis of the capsid proteins is one of the key regulators of B19 tissue tropism, and governs the ability of AMDV to remain persistent in infected host animals. B19 permissive infection is characterized by a switch to increased capsid production, while persistence of AMDV infection in animal systems is characterized by, and likely requires, controlled low levels of capsid protein synthesis. All other parvoviruses have an internal promoter that regulates production of capsid-encoding mRNA during the synthesis phase of viral replication, however, this mechanism is not available to B19 and AMDV. Control of expression of the AMDV and B19 capsid-coding genes by post-transcriptional mechanisms is the focus of this application. Both viruses have an efficient polyA site in the center of the genome. Use of this site precludes inclusion of the capsid coding ORFs into mRNA, and an understanding of how the choice is made to either polyadenylate, or read-through this site, is the topic of Specific Aim 2 and 3. We propose to identify both the cis sequence and trans-factors that control these events, the mechanisms that govern their selective use, and for B19, exam how this choice is differently made in cells permissive or nonpermissive for replication. We have recently discovered that a single spliced AMDV mRNA encodes both capsid proteins VP1 and VP1, as well as an essential small non-structural protein NS2 at its 5'end. Thus, in addition to alternative splicing and polyadenylation, features of alternative translation initiation are critical for governing the proper expression of the AMDV capsid proteins.
In Specific Aim 1, we will characterize how the cis-element in the NS2 gene affects translation of the capsid proteins, both genetically and biochemically. How these post-transcriptional processes generate appropriate levels of the capsid proteins from the single pre-mRNA molecules encoded by AMDV and B19, is critical to our understanding of parvovirus gene expression, and the biology of parvovirus infection. However, in addition, these viral systems provide very tractable models with which to learn much about these basic cellular mechanisms in general.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI070723-05
Application #
8120571
Study Section
Virology - B Study Section (VIRB)
Program Officer
Park, Eun-Chung
Project Start
2007-08-01
Project End
2013-05-09
Budget Start
2011-08-01
Budget End
2013-05-09
Support Year
5
Fiscal Year
2011
Total Cost
$282,675
Indirect Cost
Name
University of Kansas
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
016060860
City
Kansas City
State
KS
Country
United States
Zip Code
66160
Pillay, Sirika; Zou, Wei; Cheng, Fang et al. (2017) AAV serotypes have distinctive interactions with domains of the cellular receptor AAVR. J Virol :
Wang, Zekun; Shen, Weiran; Cheng, Fang et al. (2017) Parvovirus Expresses a Small Noncoding RNA That Plays an Essential Role in Virus Replication. J Virol 91:
Qiu, Jianming; Söderlund-Venermo, Maria; Young, Neal S (2017) Human Parvoviruses. Clin Microbiol Rev 30:43-113
Deng, Xuefeng; Xu, Peng; Zou, Wei et al. (2017) DNA Damage Signaling Is Required for Replication of Human Bocavirus 1 DNA in Dividing HEK293 Cells. J Virol 91:
Deng, Xuefeng; Zou, Wei; Xiong, Min et al. (2017) Human Parvovirus Infection of Human Airway Epithelia Induces Pyroptotic Cell Death by Inhibiting Apoptosis. J Virol 91:
Wang, Zekun; Deng, Xuefeng; Zou, Wei et al. (2017) Human Bocavirus 1 Is a Novel Helper for Adeno-Associated Virus Replication. J Virol :
Shen, Weiran; Deng, Xuefeng; Zou, Wei et al. (2016) Analysis of cis and trans Requirements for DNA Replication at the Right-End Hairpin of the Human Bocavirus 1 Genome. J Virol 90:7761-77
Deng, Xuefeng; Yan, Ziying; Cheng, Fang et al. (2016) Replication of an Autonomous Human Parvovirus in Non-dividing Human Airway Epithelium Is Facilitated through the DNA Damage and Repair Pathways. PLoS Pathog 12:e1005399
Luo, Yong; Qiu, Jianming (2015) Human parvovirus B19: a mechanistic overview of infection and DNA replication. Future Virol 10:155-167
Shen, Weiran; Deng, Xuefeng; Zou, Wei et al. (2015) Identification and Functional Analysis of Novel Nonstructural Proteins of Human Bocavirus 1. J Virol 89:10097-109

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